Hervé Bernard1, Paul J Turner2, Sandrine Ah-Leung1, Monica Ruiz-Garcia2, Elizabeth Naomi Clare Mills3, Karine Adel-Patient1. 1. CEA, INRAE, Département Médicaments et Technologies pour la Santé (DMTS)/ Service de Pharmacologie et d'Immunoanalyse, Université Paris-Saclay, Gif-sur-Yvette, France. 2. Section of Inflammation, Repair & Development, National Heart & Lung Institute, Imperial College London, London, UK. 3. Division of Infection, Immunity and Respiratory Medicine, School of Biological Sciences and Manchester Institute of Biotechnology, Manchester Academic Health Sciences Centre, University of Manchester, Manchester, UK.
Abstract
BACKGROUND: Bioaccessibility of food allergens may be a key determinant of allergic reactions. OBJECTIVE: To develop a protocol allowing the detection of the major peanut allergen, Ara h 6, in the bloodstream following ingestion of low amounts of peanut and to compare Ara h 6 bioaccessibility by food matrix. We further assessed for differences in absorption in healthy versus peanut-allergic volunteers. METHODS: A blood pretreatment combining acidic shock and thermal treatment was developed. This protocol was then applied to blood samples collected from human volunteers (n = 6, healthy controls; n = 14, peanut-allergic patients) at various time-points following ingestion of increasing levels of peanut incurred in different food matrices (cookies, peanut butter and chocolate dessert). Immunodetection was performed using an in-house immunoassay. RESULTS: An original pretreatment protocol was optimized, resulting in irreversible dissociation of human antibodies-Ara h 6 immune complex, thus rendering Ara h 6 accessible for its immunodetection. Ara h 6 was detected in samples from all volunteers following ingestion of 300-1000 mg peanut protein, although variations in the kinetics of passage were observed between individuals and matrices. Interestingly, in peanut-allergic subjects, Ara h 6 could be detected following ingestion of lower doses and at higher concentrations than in non-allergic volunteers. CONCLUSIONS AND CLINICAL RELEVANCE: The kinetics and intensity of Ara h 6 passage in bloodstream depend on both individual and food matrix. Peanut-allergic patients appear to demonstrate higher absorption rate, the clinical significance of which warrants further evaluation.
BACKGROUND: Bioaccessibility of food allergens may be a key determinant of allergic reactions. OBJECTIVE: To develop a protocol allowing the detection of the major peanut allergen, Ara h 6, in the bloodstream following ingestion of low amounts of peanut and to compare Ara h 6 bioaccessibility by food matrix. We further assessed for differences in absorption in healthy versus peanut-allergic volunteers. METHODS: A blood pretreatment combining acidic shock and thermal treatment was developed. This protocol was then applied to blood samples collected from human volunteers (n = 6, healthy controls; n = 14, peanut-allergicpatients) at various time-points following ingestion of increasing levels of peanut incurred in different food matrices (cookies, peanut butter and chocolate dessert). Immunodetection was performed using an in-house immunoassay. RESULTS: An original pretreatment protocol was optimized, resulting in irreversible dissociation of human antibodies-Ara h 6 immune complex, thus rendering Ara h 6 accessible for its immunodetection. Ara h 6 was detected in samples from all volunteers following ingestion of 300-1000 mg peanut protein, although variations in the kinetics of passage were observed between individuals and matrices. Interestingly, in peanut-allergic subjects, Ara h 6 could be detected following ingestion of lower doses and at higher concentrations than in non-allergic volunteers. CONCLUSIONS AND CLINICAL RELEVANCE: The kinetics and intensity of Ara h 6 passage in bloodstream depend on both individual and food matrix. Peanut-allergicpatients appear to demonstrate higher absorption rate, the clinical significance of which warrants further evaluation.
Authors: Rebecca Czolk; Julia Klueber; Martin Sørensen; Paul Wilmes; Françoise Codreanu-Morel; Per Stahl Skov; Christiane Hilger; Carsten Bindslev-Jensen; Markus Ollert; Annette Kuehn Journal: Front Immunol Date: 2021-01-28 Impact factor: 7.561
Authors: Paul J Turner; Stefania Arasi; Barbara Ballmer-Weber; Alessia Baseggio Conrado; Antoine Deschildre; Jennifer Gerdts; Susanne Halken; Antonella Muraro; Nandinee Patel; Ronald Van Ree; Debra de Silva; Margitta Worm; Torsten Zuberbier; Graham Roberts Journal: Allergy Date: 2022-04-28 Impact factor: 14.710